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Methods Mol Biol. 2012;884:319-34. doi: 10.1007/978-1-61779-848-1_23.

Retinal transcriptome profiling by directional next-generation sequencing using 100 ng of total RNA.

Author information

1
Neurobiology Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD, USA.

Abstract

RNA expression profiles produced by next-generation sequencing (NGS) technology (RNA-seq) allow comprehensive investigation of transcribed sequences within a cell or tissue. RNA-seq is rapidly becoming more cost-effective for transcriptome profiling. However, its usage will expand dramatically if one starts with low amount of RNA and obtains transcript directionality during the analysis. Here, we describe a detailed protocol for the creation of a directional RNA-seq library from 100 ng of starting total RNA.

PMID:
22688717
DOI:
10.1007/978-1-61779-848-1_23
[Indexed for MEDLINE]

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